• 한국자원식물학회:학술대회논문집
• /
• 한국자원식물학회 2003년도 심포지엄
• /
• pp.50-58
• /
• 2003

Isolation and culture of leaf protoplasts from two chilli cultivars (Capsicum annuum var. accumnatum and Bird chilli) were developed to enhance selection process in the somatic hybridization programmes. In order to isolate the protoplasts from leaves of these two chilli cultivars different incubation periods (3, 5 and 10 hours) were tested with combinations of enzyme mixtures containing cellulase and macerozyme. Leaves were incubated on three enzyme mixtures (2% cellulase + 0.4% macerozyme, 1% cellulase + 0.2% macerozyme and 0.5% cellulase + 0.1 % macerozyme in 13% mannitol) at 251oC in the dark. Three hours of incubation using 2% cellulase and 0.4% macerozyme was the best for the protoplast isolation of both chilli cultivars tested. The yield was 5 ${\times}$ 108protoplasts/ml/ g leaf tissue in both chilli varieties. It was found that in the mixed nurse method using Nagata and Takebe (NT) medium supplemented with 1.0mg/12,4-D, NAA and BAP with 0.5M mannitol and 1.2% Sea Plaque agarose is the best medium for protoplast culture. Protoplasts of Capsicum annum var. accumnatum were alive for 14 days forming cell walls and initiating cell division.

• Journal of Ginseng Research
• /
• 제25권3호
• /
• pp.136-140
• /
• 2001

배양온도 및 배지의 pH가 인삼 뿌리썩음병균 Cylindrocarpon destructans 후막포자의 발아에 미치는 영향을 배지별로 조사한 결과는 다음과 같다. 배양온도별 후막포자 발아율은 Czap다 solution agar(CSA) 배지가 potato dextrose agar(PDA) 보다 높았다. CSA 배지의 경우에는 배양온도 5~3$0^{\circ}C$에서 후막포자의 발아가 가능하였으며 15~$25^{\circ}C$에서 발아율은 53.2~62.7%로서 발아적정 온도범위로 측정되었다. 후막포자 발아 최적 배양온도는 2$0^{\circ}C$로 발아율은 62.7%이었고, 3$0^{\circ}C$에서는 6.9%의 매우 저조한 발아율을 보였다. PDA배지에서는 5~$25^{\circ}C$에서 후막포자가 발아 가능하였고, 3$0^{\circ}C$에서는 발아가 관찰되지 않았다. 10~2$0^{\circ}C$에서 발아율이 43.6~47.9%로서 발아적정 온도범위로 측정되었다. 후막포자 발아최적 배양온도는 2$0^{\circ}C$로 발아율은 47.9%이었다. C. destructans 후막포자는 CSA 배지의 pH 5.2~8.1 조사범위 전체에서 발아되었으며 발아적정 pH 범위는 pH 6.4~8.1이었다. PDA배지의 경우는 pH 5.2~7.4의 조사범위 전체에서 발아가 확인되었으며 발아적정 pH 범위는 pH 5.2~6.0 이었다. C. destructans 후막포자를 21일간 배양한 후 생성된 균총의 색상을 배지 pH별로 조사한 결과, CSA 배지의 경우 pH 5.2~6.0 범위에서 연한 갈색, pH 6.4~8.1 에서 백색의 균총을 형성한 반면, PDA 배지에서는 pH 5.2~7.1에서 C. destructans의 전형적인 암갈색 균총을 형성하였고 pH 7.2에서는 갈색균총을 형성하였다.

• KSBB Journal
• /
• 제24권6호
• /
• pp.556-562
• /
• 2009

본 연구는 TNT 분해능이 우수한 세균인 S. maltophilia OK-5를 이용하여 TNT 함유 폐수인 pink water의 미생물학적 처리 가능성에 대한 연구를 하였다. Pink water에 함유된 TNT 제거를 위해 S. maltophilia OK-5를 교반탱크 반응조에서 배양한 결과 pink water 내에 존재하는 100 mg/L의 TNT를 배양 6일 만에 완전 분해하였다. Hydride-Meisenheimer complex에서 유래하는 진한 적갈색은 배양기간 내에 증가하였으며, 이를 정량적으로 확인하였다. 본 연구에서 pink water에 잔류하는 TNT 뿐만 아니라 2,4-dinitrotoluene, 2,6-dinitrotoluene, 2,4-dinitro-6-hydroxytoluene 등의 대사산물도 HPLC 분석방법으로 측정하였으며, GC-MS를 사용하여 확인하였다. 또한 pink water에서 배양된 S. maltophilia OK-5에서 발현되는 nitroreductase (pnrB)의 유전자 발현 정량을 real time PCR로 측정하였다. 그 결과 배양 5일째 pnrB copy 수가 $10^3$ 이상 증가하는 것을 확인하였다.

• 식물조직배양학회지
• /
• 제25권5호
• /
• pp.347-351
• /
• 1998

딸기 '수홍'의 기내배양된 잎과 탁엽 절편체로부터 기관형성을 통한 식물체 재생 방법을 확립하고자 실험을 수행하였다. 엽절편체 배양에서는 생장조절제 처리에 따라 NAA 첨가시에는 뿌리 재생, BA 첨가시에는 신초 재생이 이루어졌으며, BA 1.0㎎/L + NAA 0.2 ㎎/L 처리구에서 재생률 31.1%, 절편체당 신초수 1.7개로 가장 양호하였다. 탁엽절편체 배양에서는 생장조절제와 광조건의 처리를 실시하였는데, 명조건에서는 생장조절제 처리에 따른 뚜렷한 경향을 발견할 수 없었으나, 암조건에서는 고농도 BA와 저농도 NAA의 혼용처리가 신초 재생에 필요함을 알 수 있었다. 탁엽절편체는 BA 2.0 ㎎/L + NAA 0.1㎎/L 처리구에서 재생률 44.4%, 절편체당 신초수 4.0개로 가장 양호한 반응을 나타내었다. 재생된 신초는 NAA 0.1㎎/L 가 첨가된 MS배지에서 발근되었으며, 이어서 인공토양(vermiculite : perlite = 1 : 1)에서 성공적으로 활착을 유도할 수 있었다.

• Journal of Crop Science and Biotechnology
• /
• 제10권2호
• /
• pp.112-116
• /
• 2007

Kinetin(KN) is an inducer of rice(Oryza sativa L.) defense/stress responses, as evidenced by the induction of inducible secondary metabolite and defense/stress protein markers in leaf. We show a novel light-dependent effect of KN-triggered defense stress responses in rice leaf. Leaf segments treated with KN(100 ${\mu}M$) show hypersensitive-like necrotic lesion formation only under continuous light illumination. Potent accumulation of two phytoalexins, sakuranetin and momilactone A(MoA) by KN that peaks at 48 h after treatment under continuous light is completely suppressed by incubation under continuous dark. Using two-dimensional gel electrophoresis we identified KN-induced changes in ribulose-1, 5-bisphosphate carboxylase/oxygenase, energy- and pathogenesis-related proteins(OsPR class 5 and 10 members) by N-terminal amino acid sequencing and mass spectrometry. These changes were light-inducible and could not be observed in the dark(and control). Present results provide a new dimension(light modulation/regulation) to our finding that KN has a potential role in the rice plant self-defense mechanism.

• Journal of Photoscience
• /
• 제9권2호
• /
• pp.427-429
• /
• 2002

We studied iron release from ferritin by irradiating the visible light, and then followed ferritin-mediated lipid peroxidation in the rod outer segment (ROS) fraction of the porcine retina. In the presence of several phosphorus compounds such as ADP and ATP, iron release from ferritin at pH 7.0 could be induced by irradiation of the visible light to the reaction mixtures. Furthermore, iron release from ferritin in the presence of ADP depended on the incubation time and the visible light irradiation. Moreover, we investigated lipid peroxidation level in the ROS fraction by two independent assay systems including the thiobarbituric acid (TBA) and ferrous oxidation/xylenol orange (FOX) methods. The visible light induced ferritin-mediated lipid peroxidation in the ROS fraction in time- and irradiance-dependent manners. In the dark condition, iron release and lipid peroxidation were not observed. Iron release from ferritin by irradiating the visible light may play an important role in the etiology of phototoxic injuries in vivo.

• The Plant Pathology Journal
• /
• 제17권4호
• /
• pp.210-215
• /
• 2001

The phenotype of Rhizoctonia solani KR-13 was randomly segregated to both melanin-producing (M+) and non-producing (M-) types through successive cultures on PDA. M+type with dark melanin showed strong pathogenicity to rice and self-anastomosis. Meanwhile, M- type with white or less-melanized mycelia showed very weak pathogenicity and non-self-anastomosis. Melanin production of R. solani was affected by incubation temperature in both M+ and M- types, but not by light treatment. The application of tricyclazole, an inhibitor of fungal melanin biosynthesis, showed no controlling effect on R. solani causing rice sheath blight. Results of this study showed that melanization of mycelia of R. solani is an important pathogenicity factor in rice.

• 한국미생물·생명공학회지
• /
• 제22권2호
• /
• pp.115-119
• /
• 1994

White mutant of Ganoderma lucidum(G4142) induced the non-basediocarpous basidiospores(NBB) from the aerial mycelia on agar media by the light illumination. Light was found to be necessary for NBB formation, but it also inhibited the growth of mycelium. The best sporulation was obtained at the periodic exposure of 16 hour light and 8 hour dark. Blue and yellow light were the most effective on sporulation, however, near UV and red light did not induce any spores. Effective light intensity for NBB bearing was about 1,000 lux as white light. Even after 16 days of culture, this strain did not form the pinhead nor chlamydospore. Optimum temperature for the mycelial growth and NBB formation were 30$\circ $C. Ganoderma lucidum G4142 exhibited the formation of stroma after five days of incubation at 30$\circ $C.

• 임산에너지
• /
• 제23권2호
• /
• pp.21-28
• /
• 2004

표고버섯 톱밥재배시 배양에 의한 중량감소율과 생장기간에 따른 에르고스테롤 정량으로 균사 생장량을 조사하였고, 배양기간과 자실체 발생기간에 따른 균체 외와 균체 내의 효소 특성을 조사하였다. 배양기간에 따른 중량 감소는 산림5호가 농기3호와 산림6호에 비해 낮은 중량감소율을 나타냈다. 에르고스테롤 정량에 의한 균사 생장량 조사 결과, 암배양기간 동안 3균주 모두 균사 생장량이 증가하였으며, 농기3호가 상대적으로 높은 증가율을 보인 반면, 산림5호는 낮은 증가율을 보였다. 농기3호와 산림6호는 명배양기간 동안에 균사 생장량이 최고치에 이르렀으며, 산림5호는 자실체 발생기간 전까지 계속 증가하였다. 배양기간과 발생기간에 따른 글체 외와 군체 내 효소 역가를 측정한 결과, 배양 초기(10일)에는 셀룰로오스와 헤미셀룰로오스 분해에 관여하는 CMCase, avicelase, xylanase, glucanase의 역가가 높았고, 배양 10일 후 리그닌 분해에 관여하는 laccase와 Mn-peroxidase의 역가가 높아지는 경향을 나타내었다.

• 미생물학회지
• /
• 제31권3호
• /
• pp.218-223
• /
• 1993

Conditions for the release and regeneration of protoplasts form Rhizopus oryzae and intergeneric protoplast fusion between Rhizopus oryzae and Aspergillus oryzae were studied. High yields of protoplast fusion between Rhizopus oryzae and Aspergillus oxyzae were studied. High yield of protoplasts from young germilings of R. oryzae were obtained by using lytic enzymes containing chitosanase (3 mg/ml), chitinase (3 mg/ml) and Novozym 234 (5 mg/ml). 0.5M glucose was used as the osmotic stabilizer and optimum pH of buffer was determined to be pH 7.5-8.0. Under these conditions, protoplasts were formed after about 3-4 hrs incubation. Approximately, 1.0%-4.9% of these protoplasts were formed after about 3-4 hrs incubation. Approximately, 1.0%-4.9% of these protoplasts regenerated on solid medium with a soft agar overlay. We have also carried out protoplasts fusion between R. oryzae and A. oryzae and have succeeded in obtaining three types of intergeneric fusants. In these experiments, 35% PEG-4000 and 10 mM CaCl$_{2}$ were used as fsogenic agents, and auxotrophic properties were used as a genetic marker to select fusants. Complementation frequency be protoplasts fusion of A. oxyzae and R. oryzae was 4.4% * 10$^{-5}$ . The fusant strains of the first type were prototrophs showing an Aspergillus type morphology with dark-yellow sporulation, those of the second type were also Apergillus type morphology but showed no sporulation. And the strains of the third type stopped growing when fusion products grown on regeneration minimal medium were transferred to fresh minimal medium. The formation of fusion products was observed by fluorescent vital stains for complementary labelling of protoplats from R. oryzae and A. oryzae. Rhodamine 6G and fluorescein diacetate wer useful complementary vital stains of Rhizopus and Aspergillus protoplasts for visualization of requency and type (dicell, multicell) of fusion.